Short bowel syndrome (SBS), a major clinical problem affecting patients of all ages, results from the functional or anatomic loss of extensive segments of small intestine. SBS has an overall 5-year survival of 70%, and in newborn infants with less than 10% of expected intestinal length, 5-year survival is only 20%. Current treatment options are inadequate and associated with severe complications and death. Patients with SBS require total parenteral nutrition (TPN) to survive. Currently, over 20,000 SBS patients per year are on home TPN in the US alone. Annual costs of home TPN exceed $300,000 per year per patient ($6 billion/year). In addition the use of TPN is associated with numerous central venous catheter-associated infectious and thrombotic complications, resulting in additional high costs. Thus, SBS represents an extremely costly, and deadly, burden to society. At present, the treatment for SBS is mainly supportive. Although small bowel transplantation is an option, the results are suboptimal, with 1-year and 4-year survival rates of 90% and 60%, respectively, and the required life-long immunosuppression causes substantial secondary complications. Novel approaches for the treatment of patients with SBS are critically needed. Our approach to this unsolved medical problem is the production of tissue-engineered intestine (TEI) using the patient's own intestinal cells combined with a synthetic nanofiber-based scaffold, resulting in a novel solution to this unmet clinical need. In this project, we will accomplish three specific aims necessary to move towards commercialization: Aim 1) Determine the optimal cell isolation and cell seeding methodology for structural formation of TEI. Aim 2) Optimize enteric nervous system (ENS) development in TEI. Aim 3) Develop an in vivo culture standard operating procedure (SOP) for development of TEI. This Phase I SBIR project will allow us to collect the pivotal data needed to scale up to a large animal model in Phase II, and to license the technology to a commercial partner in Phase III for commercialization.